JPH0755134B2 - Method for producing red pigment by safflower cultured cells - Google Patents

Description

TECHNICAL FIELD The present invention relates to a method for producing a red pigment by cultured safflower cells.

[Conventional technology]

In recent years, it has been desired to use natural dyes, which are safer than synthetic dyes, as food coloring agents. Natural pigments are produced by being extracted from plants, etc., but they require vast land, time, and labor to cultivate plants, and their harvest depends on the weather, soil, etc. and maintains stable quality. Was difficult.

[Problems to be Solved by the Invention]

An object of the present invention is to solve the above-mentioned problems of the prior art and to provide a method for producing a red pigment from cultured safflower cells, which enables stable and constant quality production of a red pigment from cultured safflower cells. .

[Means for Solving the Problems]

The present invention provides a method for producing a red pigment from safflower cultured cells, wherein safflower cultured cells are treated with phosphate ions 23
The present invention relates to a method for producing a red pigment by safflower cultured cells, which comprises performing growth culture in a medium containing 8 mg / or more and magnesium ion 73 mg / or more, and then performing pigment production culture.

The safflower cultured cells used in the present invention are obtained by liquid-culturing callus derived from safflower tissue. The safflower tissue used for the induction of Qasuru includes seeds, cotyledons of safflower seedlings, hypocotyls, roots, mature safflower leaves, stems, flowers,
There is no particular limitation on any part of the plant such as the root. After inducing callus under appropriate conditions, these tissues are subcultured using a liquid medium or agar medium until the properties of the callus are stabilized.

The medium for culturing safflower cultured cells used in the present invention is not particularly limited, for example, known Murashige-Skoog medium (1962), Linsmaier-Skoog medium (1965), White medium (1963), Gamborg B-5 medium. (1968), Nitsch medium (1951), Heller medium (1953), Schenk-Hildebrandt
Medium (1972), Nitsch-Nitsch medium (1967), Kohlenba
Ch-Schmidt medium (1975), Knop medium (1865) or modified medium thereof is used.

In the present invention, after carrying out a growth culture using the above-mentioned medium, a pigment production culture is carried out.In the growth culture, the phosphate ion concentration is 238 mg / or more, preferably 475 mg / or more, and the magnesium ion concentration is It is necessary to use a culture medium of 73 mg / or more, preferably 110 mg / or more. If the concentration of phosphate ion and magnesium ion is less than the above concentration, dye production cannot be improved. Further, in the pigment production culture of safflower red pigment, in the above-mentioned medium, sucrose, glucose, raffinose, fructose, pentose, maltose and the like as a carbon source,
Cytokinin or auxin is added as a plant hormone. Examples of cytokinins include zeatin, dihydrozeatin, ribosylzeatin, 6-benzyladenine (BA), and kinetin, and auxins include indole-3-acetic acid (IAA), 1-naphthaleneacetic acid (NAA), and 2,4-dichloro. Phenoxyacetic acid (2,4-
D), 2,4,5-Trichlorophenoxyacetic acid (2,4,5-T)
Etc.

The culture conditions are not particularly limited, but shaking or rotary culture at a temperature of 25 ° C. in the dark is preferable. In addition, it is preferable that the growth culture is usually carried out for about 7 to 14 days, and the dye production culture usually produces the red pigment in about 7 to 14 days.

Examples Hereinafter, the present invention will be described in detail with reference to Examples.

〔Example〕

<Culture of safflower cultured cells> Murashige-containing 10 ⁻⁵ M NAA and 10 ⁻⁶ M kinetin
Safflower cotyledon slices were plated on Skoog agar to induce callus, which was then treated with 10 ^-5 M NAA and 10
Suspension culture was performed using a Murashige-Skoog liquid medium containing ^-6 M kinetin, and the cells were subcultured in the same medium for about 1 year to obtain cell clumps.

Example 1 and Comparative Example 1 A Murashige-Skoog medium (Comparative Example 1) having the composition shown in Table 1 and a medium (Example 1) prepared by doubling each concentration of the main inorganic salt of this medium were prepared at 100 rpm. Put 30 ml each in a 100 ml flask on a rotating rotary shaker,
Inoculate 0.4 g of safflower cultured cells into this, and in the dark at ambient temperature
Proliferation culture was performed at 25 ° C for 10 days.

0.25 g of the obtained cells was inoculated into the dye-producing medium shown in Table 2 and cultured in the dark at a temperature of 25 ° C. for 10 days, and the amount of the red dye produced was measured. The measurement was performed by measuring a wavelength of 215 nm with a spectrophotometer. As a result, the production amount of the red pigment in Example 1 was 3.375 mg /
It was found that the production amount was improved by 1.35 times as compared with the production amount of Comparative Example 1 of 2.5 mg /.

Example 2 and Comparative Example 2 CaCl ₂ · 2H ₂ O, KH ₂ PO ₄ and MgSO ₄ · of the medium shown in Table 1
Proliferation culture and pigment production were carried out in the same manner as in Example 1 except that a medium adjusted so that the concentration of 7H ₂ O was doubled was used, and the production amount of red pigment was measured. Further, the same safflower cultured cells used at this time were used to perform pigment production in the same manner as in Comparative Example 1, and the production amount was measured (Comparative Example 2).

As a result, the dye production amount of Example 2 was 3.013 mg /, the dye production amount of Comparative Example 2 was 1.8 mg /, and it was found that the production amount in Example 2 was improved 1.67 times as compared with Comparative Example 2. .

Examples 3 to 11 and Comparative Example 3 The media shown in Table 1 had three times the concentration of MgSO ₄ .7H ₂ O and CaCl 2
Proliferation culture and pigment production were carried out in the same manner as in Example 1 except that the medium in which the concentrations of H ₂ O and KH ₂ PO ₄ were adjusted to the magnifications shown in Table 3 were used, respectively, and the production amount of red pigment was increased. Was measured. The results are shown in Table 3. Further, the same safflower cultured cells used at this time were used to perform pigment production in the same manner as in Comparative Example 1, and the production amount was measured. As a result, the pigment production amount was 1.67 mg / (Comparative Example 3). .

From Table 3, regardless of the concentration of CaCl ₂ · 2H ₂ O in the medium, if the concentration of MgSO _{4 in} the medium is 3 times and the concentration of KH ₂ PO ₄ is 2 times or more, the dye production amount is It can be seen that It was also found that increasing the concentration of KH ₂ PO ₄ more than four times without increasing the amount of CaCl · 2H ₂ O further improved the productivity.

〔The invention's effect〕

According to the present invention, after growing and culturing in a medium containing a phosphate ion and magnesium ion at a certain concentration or more, by producing a pigment, it is possible to stably and productively produce a red pigment from safflower cultured cells. it can.

Claims (1)

[Claims] 1. A method for producing a red pigment from safflower cultured cells, wherein the safflower cultured cells are treated with phosphate ions.
A method for producing a red pigment by safflower cultured cells, which comprises performing growth culture in a medium containing 238 mg / or more and magnesium ion 73 mg / or more, and then performing pigment production culture.